Exploring the conformational landscape and stability of Aurora A using ion-mobility mass spectrometry and molecular modelling

Tomlinson, Lauren, Batchelor, Matthew ORCID: 0000-0001-6338-5698, Sarsby, Joscelyn, Byrne, Dominic, Brownridge, Philip ORCID: 0000-0003-0105-6594, Bayliss, Richard, Eyers, Patrick ORCID: 0000-0002-9220-2966 and Eyers, Claire ORCID: 0000-0002-3223-5926 (2021) Exploring the conformational landscape and stability of Aurora A using ion-mobility mass spectrometry and molecular modelling. [Preprint]

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Official URL: http://dx.doi.org/10.1101/2021.08.30.458190

Abstract

<h4>ABSTRACT</h4> Protein kinase inhibitors are proving highly effective in helping treat a number of non-communicable diseases driven by aberrant kinase signaling. They are also extremely valuable as chemical tools to help delineate cellular roles of kinase signaling complexes. The binding of small molecule inhibitors induces conformational effects on kinase dynamics; evaluating the effect of such interactions can assist in developing specific inhibitors and is deemed imperative to understand both inhibition and resistance mechanisms. Using gas-phase ion mobility-mass spectrometry (IM-MS) we characterized changes in the conformational landscape and stability of the protein kinase Aurora A (Aur A) driven by binding of the physiological activator TPX2 or small molecule inhibition. Aided by molecular modeling, we establish three major conformations: one highly-populated compact conformer similar to that observed in most crystal structures, a second highly-populated conformer possessing a more open structure that is infrequently found in crystal structures, and an additional low-abundance conformer not currently represented in the protein databank. Comparison of active (phosphorylated) and inactive (non-phosphorylated) forms of Aur A revealed that the active enzyme has different conformer weightings and is less stable than the inactive enzyme. Notably, inhibitor binding shifts conformer balance towards the more compact configurations adopted by the unbound enzyme, with both IM-MS and modelling revealing inhibitor-mediated stabilisation of active Aur A. These data highlight the power of IM-MS in combination with molecular dynamics simulations to probe and compare protein kinase structural dynamics that arise due to differences in activity and as a result of compound binding.

Item Type:	Preprint
Uncontrolled Keywords:	5.1 Pharmaceuticals, 5 Development of treatments and therapeutic interventions, 3 Good Health and Well Being
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology Faculty of Health and Life Sciences > Tech, Infrastructure and Environmental Directorate
Depositing User:	Symplectic Admin
Date Deposited:	26 Oct 2021 07:57
Last Modified:	14 Mar 2024 17:48
DOI:	10.1101/2021.08.30.458190
Related URLs:	Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3141623